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Heavy Light Chain Immunoprecipitation Problems

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Old 04-30-2008, 04:43 PM
IoDiNE IoDiNE is offline
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Question Heavy Light Chain Immunoprecipitation Problems

Hello,
i have a question for IP and antibody heavy and light chain detection.


I performed an Immunoprecipitation experiment but when i ran the SDS-PAGE gel, i loaded the rabbit antibody used for ip in a control lane however
doing the western blot with Goat anti-rabbit secondary antibody only detected a very strong 50kDa band in the lane!


Did it mean that the antibody is not completely denatured?

Should i expect another 20-30 kDa band on the gel corresponding to the antibody light chain?

I also boiled the antibody in loading buffer for 5 min so shouldnt this be ok?

Thanks again in advance
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